Rapid Analysis of Cyanide Based on a Ratiometric Fluorescent Probe Using Gold Nanoclusters-Fluorescein

doi:10.12116/j.issn.1004-5619.2025.350403

Abstract

Abstract:

Objective To establish a rapid analysis method for cyanide based on a ratiometric fluorescent probe, providing a quantitative strategy for on-site visual and rapid detection of cyanide. Methods A dual-emission ratiometric fluorescent probe （AuNCs-FL） was constructed by using bovine serum albumin （BSA）-stabilized gold nanoclusters （AuNCs, fluorescence emission at 660 nm） as the responsive signal unit and fluorescein （FL, emission at 515 nm） as the internal reference. Results The etching effect of cyanide on AuNCs resulted in fluorescence quenching at 660 nm, while the fluorescence intensity of FL at 515 nm remained unchanged, enabling a rapid response analysis of cyanide shift from red to green fluorescence. The developed probe enabled rapid analysis of cyanide within 3 min, with a limit of detection （LOD） of 3.4 mg/L and a visual detection range of 10-100 mg/L. Conclusion The AuNCs-FL fluorescent probe is structurally simple, low-cost, and easy to operate, delivering rapid and accurate results. It also avoids the interference from sulfides encountered in commercial cyanide test kits, making it suitable for the on-site rapid detection of suspected powder samples in cyanide poisoning cases.

Key words: forensic medicine, toxicological analysis, gold nanoclusters （AuNCs）, fluorescein, cyanide, ratiometric fluorescent probe, rapid analysis, portable device

CLC Number:

Tai-shen HE, Zhong-jiang LÜ, Yi-ming SUN, Yu-yang LI, Yi YE, Yao LIN, Lin-chuan LIAO. Rapid Analysis of Cyanide Based on a Ratiometric Fluorescent Probe Using Gold Nanoclusters-Fluorescein[J]. Journal of Forensic Medicine, 2025, 41(4): 340-347.

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URL: http://www.fyxzz.cn/EN/10.12116/j.issn.1004-5619.2025.350403

http://www.fyxzz.cn/EN/Y2025/V41/I4/340

Figures/Tables 10

Fig. 1 Photographs of the self-made portable fluorescence reading device

Fig. 2 Spectral characterization images of AuNCs

Fig. 3 Spectral characterization images of the constructed ratiometric fluorescent probe

Fig. 4 Optimization of reaction conditions for the AuNCs-FL probe and cyanide

Fig. 5 Fluorescence images of single AuNCs， FL，as well as AuNCs-FL after reacting with cyanideat a series of mass concentrations

Fig. 6 Standard curve of cyanide massconcentration versus G/R ratio

Tab. 1 Performance comparison between this method and other fluorescent analysis methods for cyanide

氰化物荧光分析方法	检出限	反应时间	便携式检测装置	数据来源
碳点-AuNCs法	0.15 μmol/L	-	无	[17]
香豆素探针法	0.22 μmol/L	-	无	[23]
氰乙烯基探针法	12.4 nmol/L	30 s	无	[24]
三重荧光探针法	45 nmol/L	>13 min	无	[25]
Benzo-Hemicyanine法	0.43 μmol/L	-	无	[26]
罗丹明B衍生物法	0.33 μmol/L	30 min	无	[27]
Hg-石墨烯量子点法	3.1 μmol/L	5 min	无	[28]
Calixarene法	0.115 μmol/L	-	无	[29]
HBT-Br-thiazolium法	1.79 μmol/L	2 min	试纸	[30]
三苯胺有机探针法	36 nmol/L	4 min	试纸	[31]
聚集诱导发光活性分子法	6.17 nmol/L	-	试纸	[32]
AuNCs-FL法	52.3 μmol/L	2 min	便携式装置（可读数）	本研究

Fig. 7 Results of performance analysis forcyanide using this method

Fig. 8 Fluorescence images of three types of samplesbefore and after the addition of cyanide

Tab. 2 Detection values and recovery rates of three types of samples spiked with cyanide

样品	氰化物加标质量浓度/（mg·L^-1）	检测值/（ $x ¯$ ±s，mg·L^-1）	回收率/%	RSD/%
白糖	30	30.7±0.6	102.3	1.2
淀粉	30	26.0±1.8	86.7	4.5
泥土	30	29.9±2.2	99.7	4.4

Tab. 2 Detection values and recovery rates of three types of samples spiked with cyanide

样品	氰化物加标质量浓度/（mg·L^-1）	检测值/（ $x ¯$ ±s，mg·L^-1）	回收率/%	RSD/%
白糖	30	30.7±0.6	102.3	1.2
淀粉	30	26.0±1.8	86.7	4.5
泥土	30	29.9±2.2	99.7	4.4

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